1.Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells with the combined induction of hepatocyte growth factor and fibroblast growth factor
Chinese Journal of Tissue Engineering Research 2009;13(45):8811-8816
BACKGROUND:Fibroblast growth factor can promote proliferation of mesenchymal stem cells (MSCs).grew along the wall.However,there are few reports on the differentiation of MSCs into hepatocytes following fibroblast growth factor induction.When mass concentration of hepatocyte growth factor was 1 μg/L,it can promote mitosis of hepatocytes,and is the strongest mitogenic agent for normal hepatocytes.OBJECTIVE:To explore the biological characteristics of human umbilical cord MSCs/n vitro and their differentiation ability to hepatocyte-like cells under the induction of chemical factors.DESIGN,TIME AND SETTING:The cytological in vitro study was conducted at the Institute of Blood,Jinan University from August 2008 to April 2009.MATERIALS:Umbilical cord was obtained from healthy full-term fetus,which was provided by the Guangzhou Huaqiao Hospital.The parturient signed informed consent.Hepatocyte growth factor and flbroblast growth factor were bought from Peprotech,USA.METHODS:The MSCs from human cord were isolated and cultured by type Ⅳ colagenase digestion+differential adherence.At the third passage,MSCs received cell surface antigen analysis and cell cycle determination to detect their ability to differentiate into adipocytes and osteoblasts.At the fifth passage,MSCs were adjusted into 5×10~9/L and assigned into 2 groups.BMSCs in the control group were incubated in DMEM/F12 containing 5% fetal bovine serum.BMSCs in the induction group were treated with above-mentioned medium supplemented with 20 μg/L hepatocyte growth factor and 10 μg/L flbroblast growth factor.MAIN OUTCOME MEASURES:The following parameters were measured:biological characteristics of human umbilical cord MSCs and differentiation of human umbilical cord MSCs into hepatocyte-like cells in vitro.RESULTS:At the third passage,MSCs derived from human cord expressed CD29,CD44,CD105,but not antigens of hematopoietic CD34,CD45,and 92.2% of them were in G_0/G_1 phase.Oil red O staining showed cytoplasm presented red granules.Alkaline phosphatase staining demonstrated that cytoplasm was black,with the differentiation ability into adipocytes and osteoblasts.Following 10 days of combined induction of hepstocyte growth factor and fibroblast growth factor,RT-PCR and Western blot results confirmed that cells expressed alpha fetoprotein and albumin.Negative expression was found in the control group.CONCLUSION:Human umbilical cord contained plenty of MSCs,with strong potential of multi-differentiation.Umbilical cord MSCs can differentiate into hepatocyte-like cells following combined induction of hepatocyte growth factor and fibroblast growth factor.
2.Comparison of status and predictors of academic burnout between Korean Nationality and Han Nationality nursing students
Jian ZHU ; Man LUO ; Anmin ZHANG ; Huan ZHANG ; Lijuan XU
Chinese Journal of Practical Nursing 2016;32(3):226-229
Objective To investigate the status and difference of learning burnout between Korean-Chinese nursing students and Chinese nursing students;and to compare the influencing factors of learning burnout between them.Methods A total of 307 nursing students in Yanbian University were recruited by convenience samphng method.Data were analyzed by SPSS 20.0,using descriptive statistics,t-test,ANOVA,correlation analysis,and linear regression analysis.Results The emotion and learning burnout of Korean-Chinese nursing students scored (3.18±0.52)points and (2.84±0.33)points,which were higher than (3.04±0.53)points and (2.69±0.36) points of Chinese nursing students (t=5.72,4.19,all P < 0.05);The significant predictors of learning burnout of Korean-Chinese nursing students were effective commitment,continuance commitment and ideality commitment;and the significant predictors of Chinese nursing students were effective commitment,continuance commitment and behavior self-efficacy.Conclusions Because of the special background of Korean-Chinese nursing students,it is suggested payed attention to improve their major commitment and learning self-efficacy,then to decrease their learning burnout.
4.The role of miR-301 in colon cancer cell proliferation and apoptosis
Zhonglai ZHANG ; Huan XIONG ; Bing ZHANG ; Peiqian ZHU
China Oncology 2013;(6):432-438
Background and purpose: The miR-224 in a variety of malignant tumors is overexpression, however, its expression and function in colon cancer are not clear. The aim of this study was to investigate the expression of miR-301 in colon cancer tissues and demonstrate the regulative effects of miR-301 ASO on the proliferation and apoptosis of colon cancer cell in vitro and in vitro. Methods:The expression of miR-301 in 120 colon cancer tissues and their adjacent tissues was detected by real-time quantitative PCR method. After transfection with miR-301ASO, the biological effects of miR-301 in SW620 cells were measured by MTT assay, the colony formation experiment, flow cytometry and the in vivo experiment. Results: The expression level of miR-301 was found to be overexpressed in 63.33% (76/120) of the colon cancer cases (P<0.05). miR-301 expression in SW620 cells (transfection with miR-301 ASO, 0.09±0.01) was significantly less than control group (0.50±0.07, P=0.00). MTT assay results showed that SW620 cells survived rate at 24, 48 and 96 h decreased greatly after transfection with miR-301ASO (P=0.00). Clone formation assay revealed that miR-301 ASO group colony formation rate (5.33%±0.74%) was significantly lower than the control group (33.33%±8.38%, P=0.00). In vivo study further confirmed that miR-301ASO could inhibit the proliferation of SW620 cells (P<0.05), and miR-301ASO group grew substantially slow compared with the negative control group (P=0.00). Flow cytometry indicated that the apoptotic index in miR-301 ASO group (15.68±1.46) was significantly higher than the control group (3.36±0.88, P=0.02). In addition, the Bcl2 mRNA and protein were significantly decreased after reduce the expression of miR-301 (P=0.00, P=0.00). Conclusion:MiR-301 was overexpressed in human colon cancer. Reduce the expression of miR-301 can effectively inhibit the growth of colon cancer cells and promote apoptosis. MiR-301 may become a new target for the regulation of gene expression in colon cancer.
5.Establishment of animal model of intestinal motility disorders after severe craniocerebral trauma
Huan FANG ; Limei ZHANG ; Suofei ZHANG ; Jingci ZHU
Chinese Journal of Trauma 2013;(2):165-169
Objective To build model of intestinal motility disorders (IMD) in mice after severe craniocerebral trauma by using modified free fall impact method so as to lay the foundation for further study on development mechanism of postinjury IMD.Methods Severe craniocerebral trauma in mice was induced by a cylindrical sleeve rod impactor.Pathological changes of cerebral and intestinal tissue in mice were observed at 1,6 hours and 1,3,7 days after severe craniocerebral trauma.Brain water content and intestinal propulsion percentage were detected in each time point as well.Results Acute inflammation and congestion of the intestine were observed in phase of cerebral hemorrhage and edema after severe craniocerebral trauma in mice.Brain water content was increased at 6 hours and reached the peak at 24 hours.Intestine propulsion percentage showed a fall at one hour and dropped to the lowest level at 6 hours.Conclusions IMD exists in mice with severe craniocerebral trauma.The device involving modified Feeney free fall impact method can successfully establish animal model of IMD in mice with severe craniocerebral trauma.
6.Screening Analysis of Neonatal Congenital Hypothyroidism and Phenylketonuria in Lianyungang Area
li-mei, ZHU ; huan-huan, ZHANG ; liang-yi, QIN ; wen-bin, ZHAO
Journal of Applied Clinical Pediatrics 2006;0(20):-
Objective To explore the incidence and distribution features of neonatal phenylketonuria(PKU) and congenital hypothyroidism(CH) in Lianyungang area.Methods The heel blood samples dried on filter paper was collected from newborns after 72 hours.As the screening indicators,PKU was detected through serum phenylalanine concentration(Phe) detection by Guthrie′s bacterial inhibition assay and CH was detected through serum thyroid stimulating hormone(TSH) levels detection by time-resolved fluorometry(TRFIA) or enzyme linked immunosorbent assay(ELISA).Results Sixty-four children with CH were diagnosed and the rate of CH was 1∶2423,41 patients were examined by thyroid gland nuclein kinescope picture,among which there were 23 patients whose thyroid glands were unusal.These patients lived in 45 towns(streets),which had no difference between men and women,city and country side.A couple of twins were screened for CH;15 cases of PKU was detected with an incidence of 1∶10 339.These patients′ parents were neither consanguineous marriage nor dominance hereditary family history.Their physical and mental development were similar to the same age children after interventive treatment.Conclusions The distribution of CH and PKU is diffuse in Lianyungang.Neonatal screening is the only valid means to find patients with CH and PKU.
7.Utilize the simplified POP-Q system in the clinical practice of staging for pelvic organ prolapse:comparative analysis with standard POP-Q system
Huan ZHANG ; Lan ZHU ; Tao XU ; Jinghe LANG
Chinese Journal of Obstetrics and Gynecology 2016;51(7):510-514
Objective To determine the association between simplified pelvic organ prolapse quantification system (S-POP-Q) and the standard pelvic organ prolapse quantification system (POP-Q) in describing pelvic organ prolapse. Methods This was an observational study. From Jan. 2010 to Jan. 2014, 256 subjects with pelvic floor disorder symptoms underwent two exams: a POP-Q exam and a S-POP-Q exam. For the S-POP-Q system, vaginal segments of the exam were defined using points Ba, Bp, C, and D. For the POP-Q system vaginal segments of the exam were defined using points Aa, Ba, Ap, Bp, C, and D. The inter-system consistency between the overall ordinal stages, the anterior vaginal wall stages, the posterior vaginal wall stages, the cervix stages, the posterior fornix or vaginal cuff stages from each two kind of exam were compared. Results The Kendall tau-b correlation coefficient for overall stage was 0.81, the Kendall tau-b correlation coefficients were 0.81, 0.81, 0.85, 0.88 for the anterior vaginal wall, for the posterior vaginal wall, for the cervix, for the posterior fornix or vaginal cuff, respectively. Conclusion There is almost perfect association between S-POP-Q and POP-Q in describing pelvic organ prolapse.
8.Effects of Chaihu Shugan San on Autonomic Behaviors and Spatial Learning- memory Abilities of Rats Treated with Stress for Different Time
Yanfang ZHU ; Jiangxia ZHOU ; Huan CHEN ; Mingjia ZHANG ; Haiqing AO
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(5):708-712
Objective To observe the effects of stress treatment for different time on autonomic behaviors and spatial learning-memory abilities of rats and to investigate the therapeutic efficacy of Chaihu Shugan San(CSS). Methods Seventy-five 8-week-old Wistar rats were randomly divided into blank control group, model control group, and low-, middle-, and high- dose CSS groups, 15 rats in each group. Chronic multi-stress model was set up. The rat body weight in various groups was measured. The autonomic behaviors of rats were observed by open field test, and the abilities of spatial learning-memory of rats were examined by Morris water maze test. Results (1)On modeling day 20, 30, the model control group had lighter body weight than the blank control group (P < 0.01); the middle-, high- dose CSS groups had higher body weight than the model control group (P < 0.05 or P < 0.01) , and the high-dose CSS group had the highest body weight, the difference being insignificant compared with the blank control group (P > 0.05). (2)Open field test results showed as follows:Compared with the blank control group, the total distance and central-area distance were shortened (P < 0.01) and the average speed was decreased(P < 0.01) in the model control group at different modeling time points, the effect being positively correlated with the modeling time; compared with the model control group, the total distance and central-area distance were prolonged(P < 0.05 or P < 0.01) and the average speed was increased (P < 0.05 or P < 0.01) in the low-, middle-, and high- dose CSS groups at different modeling time points, and the therapeutic efficacy of high- dose CSS group was approach to the blank control group. (3)Morris water maze test results showed as follows: Compared with blank control group, the average latent period was prolonged and the times of platform crossing were decreased in the model control group (P < 0.01); compared with the model control group, the average potential time was shortened and the times of platform crossing were increased in the middle- and high-dose CSS groups (P<0.01), the efficacy being not significantly different from that of the blank control group (P > 0.05). Conclusion The chronic stress may decrease the excitability, reduce autonomic behaviors, and injure spatial learning and memory abilities of rats, the effect being positively correlated with the modeling time. And CSS has an effect on improving the above indexes, in particular the middle-and high-dose of CSS.
9.Expression and significance of HER2 in 108 cases of colon carcinoma
Huan XIONG ; Zhonglai ZHANG ; Bin LAI ; Hongliang LUO ; Peiqian ZHU
Chongqing Medicine 2014;(2):185-187,190
Objective To detect the expression of HER2 in clinical colon carcinoma tissue ,to investigate its correlation with the clinicopathological characteristics and to analyze its influence on the proliferation and cell cycle in colon carcinoma cell lines .Methods 108 specimens of colon carcinoma and corresponding paracancerous tissues were collected .The hybridization in situ and real-time quantitative polymerase chain reaction were used to detect the HER 2 expression in those specimens .The relationship between HER2 expression and the clinicopathologic features was analyzed .The expression of HER2 in colon carcinoma cells(SW480 and Lo-Vo) was reduced by using the antisense technology .The MTT assay and the flow cytometry were used to investigate the HER2 in-fluences on the cell proliferation and cell cycles .Results The hybridization in situ results showed that the HER2 positive expres-sion rate was 66 .67% in colon carcinoma and 10 .19% in the paracancerous tissues ,the difference between them was statistically significant(P<0 .05) .The real-time fluorescent quantitative PCR results further showed that HER2 was found to be overexpressed in 61 .11% of the colon carcinoma tissue(P<0 .05);the expression of HER2 was gradually increased with the progress of colon cancer .(P<0 .05);the expression of HER2 in the colon tissue with lymph node metastasis was also significantly higher than that without lymph node metastasis in colon carcinoma (P<0 .05);siRNA-HER2 could significantly reduce the expression of HER2 in colon cancer cell lines(SW480 and LoVo) ,the growth of colon carcinoma cell lines was also significantly inhibited and the propor-tion of cells in G0/G1 phase was increased ,while the proportion of cells in S phase and G2/M phase was decreased .Conclusion HER2 is closely related with the occurrence and development of colon carcinoma ,its mechanism could regulate the grow th of colon carcinoma cells via mediating the transition of G1/S phase ,which may provide a new target for the treatment of colon carcinoma .
10.THREE-DIMENSIONAL MORPHOLOGY OF HEPATOCELLULAR CARCINOMA——THE APPLICATION OF CONFOCAL LASER SCANNING MICROSCOPY IN TUMOR PATHOLOGY
Wanghai ZHANG ; Shineng ZHU ; Shilun LU ; Yalin HUAN ; Peng ZHAO
China Oncology 1998;0(04):-
PURPOSE To investigate the application of CLSM in tumor pathology with the three dimensional reconstruction by confocal laser scanning microscopy in routine pathologic specimens of HCC, METHODS 30 ?m thick sections were cut from the paraffin-embedded tissues of HCC. hyperplasia and normal liver, stained with the DNA fluorescent probe YOYO-1 iodide and examined by CLSM to collect optical sections and 3D reconstructed images. RESULTS HCC displayed chaotic arrangement of carcinoma cell nuclei, marked pleomorphism. indented and irregular nuclear surface, and irregular and coarse chromatin texture. CONCLUSION The serial optical tomograms of CLSM can be used to create 3D reconstruction of cancer cells. Such 3D impressions might be helpful or even essential in arriving at an exact diagnosis